积雪草苷多功能型水凝胶支架制备及对细胞黏附和三维培养的应用价值

Preparation of multifunctional hydrogel scaffolds containing Centella asiatica and its application value in cell adhesion and 3D cell culture

目的 利用积雪草苷中药单体对海藻酸钠水凝胶支架材料进行改性研究,提高其机械性能,为细胞提供更优的生存环境。方法以甲基丙烯酸酐化海藻酸钠为支架材料、N-乙烯基吡咯烷酮(N-Vinyl-2-pyrrolidone,NVP)和中药单体为造孔剂构建三维载细胞多孔复合水凝胶支架。通过表征其基本理化性质,并从机械特性、生物相容性、细胞共培养等方面评价其应用于细胞黏附和三维培养的应用价值。结果当甲基丙烯酸酐化海藻酸钠与NVP为最适比例,且积雪草苷浓度为6 mg/mL时,测得水凝胶成胶后性能最稳定、交联程度最高。与空白组相比,其溶胀率最高为45%,质量损失率最高达61.5%,对凝胶的降解有明显的影响。初步的细胞相容性研究发现,该复合水凝胶支架无细胞毒性,可实现肾小管上皮细胞的增殖,黏附于表面的细胞呈取向性生长,形成单细胞层,水凝胶支架内的细胞呈三维生长。结论 本实验所获中药单体型水凝胶支架在促进细胞黏附、增长的同时还有利于细胞迁移,可为中药天然产物分子组装成绿色凝胶技术提供新思路,对加快天然支架材料在组织工程领域的实际应用具有极其重要的意义。

Objective The modification of sodium alginate hydrogel scaffold materials using Centella asiatica herbal monomers was studied to improve their mechanical properties and provide a better living environment for cells.Methods A three-dimensional cell-carrying porous composite hydrogel scaffold was constructed using sodium methacrylate-anhydride alginate as the scaffold material,N-Vinyl-2-pyrrolidone(NVP)and traditional Chinese medicine monomer as the pore-forming agent.Characterize its basic physicochemical properties and evaluate its value for application to cell adhesion and 3D culture in terms of mechanical properties,biocompatibility,and cell co-culture.Results The most stable performance and the highest degree of cross-linking of the hydrogel after gel formation was measured when the optimal ratio of sodium methacrylate-anhydride alginate to NVP and the concentration of Centella asiatica was 6 mg/mL.The highest swelling rate was 45%and the highest mass loss rate was 61.5%compared with the blank group,which had a significant effect on the degradation of the gel.Preliminary cytocompatibility studies revealed that the composite hydrogel scaffold was non-cytotoxic and enabled proliferation of renal tubular epithelial cells,with cells adhering to the surface growing in an oriented manner to form a single cell layer and cells within the hydrogel scaffold growing in three dimensions.Conclusion The obtained herbal monomer-type hydrogel scaffold promotes cell adhesion and growth while also facilitating cell migration,which could provide new ideas for the assembly of herbal natural promoted molecules into green gel technology,and was of great research significance in accelerating the practical application of natural scaffold materials in the field of tissue engineering.